青钱柳多糖对大鼠脂代谢相关基因DNA甲基化及表达影响的研究

发布时间：2018-03-27 06:27

本文选题：青钱柳多糖　切入点：高脂血症　出处：《江西农业大学》2016年硕士论文

【摘要】：为研究青钱柳多糖对高脂血症大鼠脂代谢相关基因DNA甲基化和mRNA表达的影响,将建模成功的高脂血症大鼠随机分为青钱柳多糖高、中、低剂量组、辛伐他汀组、高脂模型组和空白对照组,每组分别灌胃青钱柳多糖水溶液400 mg/kg·体重·每天、200 mg/kg·体重·每天、100 mg/kg·体重·每天、4 mg/kg·体重·每天辛伐他汀水溶液,另两组灌胃等量无菌水,每组16只,连续喂养8周。实验结束后采用亚硫酸盐测序法(Bisulfite Sequencing PCR,BSP)和Q-PCR测定大鼠肝脏和脂肪组织中瘦素(LEPTIN)、黑素皮质素受体-4(MC4R)、微粒体甘油三酯转运蛋白(MTTP)、阿片-促黑素细胞皮质素原(POMC)、过氧化物酶体增殖体激活受体-γ(PPARγ)五个基因启动子区域DNA甲基化水平、mRNA表达情况、甲基转移酶活性以及和它们之间的相关性,以探究青钱柳多糖在脂代谢方面的分子机理。1)试验结果表明,青钱柳多糖(CPP)可以显著降低高脂血症大鼠肝脏组织中LEPTIN、MC4R、MTTP、POMC、PPARγ五个基因启动子区DNA甲基化水平(P0.05或P0.01);青钱柳多糖可以显著降低脂肪组织中LEPTIN基因DNA甲基化水平(P0.05),显著提高POMC基因DNA甲基化水平(P0.05),对MC4R、MTTP、PPARγ脂肪组织中的DNA甲基化水平无显著影响。2)Q-PCR结果表明,青钱柳多糖(CPP)能够显著下调高脂血症大鼠肝脏和脂肪组织中LEPTIN、MC4R、MTTP、PPARγ等基因的mRNA表达量(P0.05或P0.01),对高脂血症大鼠肝脏和脂肪中POMC基因mRNA的表达量无显著的影响。3)青钱柳多糖可显著抑制高脂血症大鼠脂肪中甲基转移酶(DNMTs)活性(P0.05),对肝脏中DNMTs活性有一定抑制作用,但效果不明显;4)相关性分析结果显示,在肝脏组织中,LEPTIN、MC4R、PPARγ、MTTP的DNA甲基化水平与mRNA表达量无显著相关性,在脂肪组织中,LEPTIN、MC4R、MTTP、POMC的DNA甲基化水平与mRNA表达量无显著相关性,PPARγ基因呈现极显著相关,在肝脏和脂肪组织中LEPTIN、MC4R、PPARγ三个基因DNA甲基化水平与甲基转移酶活性呈显著正相关,MTTP、POMC基因DNA甲基化水平与甲基转移酶活性正无显著相关性。
[Abstract]:In order to study the effect of Polysaccharide on DNA methylation and mRNA expression of lipid metabolism-related genes in hyperlipidemic rats, the established hyperlipidemic rats were randomly divided into three groups: high, medium, low dose and simvastatin groups. In the hyperlipidemia model group and the blank control group, the rats in each group were administrated with 400 mg/kg / day water of Cyclamen Polysaccharide (400 mg/kg / day) and 100 mg/kg / day (#number0# mg/kg / day) with simvastatin solution (4 mg/kg / day), and the other two groups were given the same amount of aseptic water by stomach. 16 in each group, After 8 weeks of continuous feeding, the rat liver and adipose tissue were determined by bisulfite Sequencing Sequencing BSPP and Q-PCR. The levels of leptin, melanin receptor -4MC-4RN, microsomal triglyceride transporter (TTP), and opioid melanocyte-stimulating cell skin were determined after 8 weeks of continuous feeding. The expression of DNA methylation level in five promoter regions of POMCs, peroxisome proliferator activated receptor-纬 PPAR- 纬, was observed. The activity of methyltransferase and its correlation with them in order to explore the molecular mechanism of lipids metabolism of Polysaccharide of Cyclocylium chinensis showed that, Polysaccharide of Salix Cycariae could significantly reduce the level of DNA methylation in the five promoter regions of LEPTINN MC4RMTTPN POMCPAR纬 in hyperlipidemic rats (P0.05 or P0.01), while Polysaccharide could significantly reduce the level of DNA methylation of LEPTIN gene (P0.05) in adipose tissue of rats with hyperlipidemia. High DNA methylation level of POMC gene (P0.05) had no significant effect on DNA methylation level in MC4RN MTTPPAR- 纬 adipose tissue. Cyclocycline Polysaccharide (CPP) could significantly down-regulate the expression of mRNA in the liver and adipose tissue of rats with hyperlipidemia by P0.01or P0.01P, but had no significant effect on the mRNA expression of POMC gene in liver and fat of hyperlipidemic rats. Glucose could significantly inhibit the activity of methyltransferase (DNMTs) in fat of hyperlipidemic rats and the activity of DNMTs in liver. The results of correlation analysis showed that there was no significant correlation between the DNA methylation level and the expression of mRNA in liver tissue. In adipose tissue, there was no significant correlation between the DNA methylation level and the expression of mRNA. In liver and adipose tissue, there was no significant positive correlation between the DNA methylation level of LEPTINN MC4RPAR- 纬 gene and the activity of methyltransferase. There was no significant positive correlation between DNA methylation level and methyltransferase activity of MTTPPOMC gene in liver and adipose tissue.
【学位授予单位】：江西农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R285.5

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